IGEM:UC Berkeley/2006/Plasmid Transformation: Difference between revisions
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2. Incubate on ice for 10 min.<br> | 2. Incubate on ice for 10 min.<br> | ||
3. Heat shock for 1.5 min at 42 C <br> | 3. Heat shock for 1.5 min at 42 C <br> | ||
4. Rescue | 4. Rescue: add 150uL of SOB media <br> | ||
5. Incubate in 37 C shaker for 1 hour<br> | 5. Incubate in 37 C shaker for 1 hour<br> | ||
6. Plate<br> | 6. Plate<br> | ||
-JL_06/14/06 | -JL_06/14/06 | ||
Revision as of 11:23, 14 June 2006
KCM-competent cell heat shock
1. create cocktail on ice
*20 uL TGI cells
*10 uL KCM buffer
*70 uL water
*1 uL plasmid
2. heat shock for 1.5 min at 42 C
3. incubate on ice for 5 min.
4. plate
-JL_06/09/06
Transformation of Ligation mix
1. Create cocktail on ice
*100 uL TGI cells
*15 uL KCM buffer
*25 uL water
*10 uL plasmid
2. Incubate on ice for 10 min.
3. Heat shock for 1.5 min at 42 C
4. Rescue: add 150uL of SOB media
5. Incubate in 37 C shaker for 1 hour
6. Plate
-JL_06/14/06
